Albumin-binding compounds that prevent nonenzymatic glycation and that may be used for treatment of glycation-related pathologies

ABSTRACT

The present invention is directed to compositions that inhibit the nonenzymatic glycation of albumin, as well as methods of using compounds that inhibit albumin glycation for the treatment of glycation-related pathologies.

RELATED APPLICATIONS

[0001] This application is a continuation-in-part application ofapplication Ser. No. 08/603,147, filed on Feb. 15, 1996, entitled“Prevention of Albumin Glycation and Complications of Diabetes withAlbumin-Binding Compounds” and of application Ser. No. 09/051,148, filedon Jan. 29, 1998, entitled “In Vivo Methods of Treatment to PreventKidney Dysfunction Using Substances that Inhibit Albumin Glycation”. Theentire disclosure is incorporated by reference herein.

TECHNICAL FIELD OF INVENTION

[0002] The present invention is directed to the discovery ofcompositions that inhibit the nonenzymatic attachment of glucose toalbumin, preventing the formation of glycated albumin. The compounds areuseful in preventing and treating disorders of the kidney and otherorgans that result from deleterious effects of glycated albumin.

BACKGROUND OF THE INVENTION

[0003] Glycated albumin, which is formed by the condensation of glucosewith reactive protein amino groups, adversely affects capillaryfunction, structure and metabolism. Experimental studies have shown thatthis glycated protein has distinct biologic effects that the nonglycatedcounterpart does not possess. These effects include facilitatedtransport across capillary filtration barriers and hyperfiltration inthe kidney, and stimulation of nitric oxide synthase and nitric oxideproduction, increased synthesis of extracellular matrix proteins, andactivation of cytokine and growth factor systems in kidney and vasculartissue. These and other biologic effects of glycated albumin have beendescribed in numerous scientific publications including KidneyInternational 42:875-881, 1992; Lab Investigation 51:27-35, 1997; KidneyInternational 45:475-484, 1994; Molecular & Cellular Biochemistry125:19-25, 1993; Molecular & Cellular Biochemistry 151:61-67, 1995;Kidney International 53:631-638, 1998.

[0004] The described biologic activities are observed withconcentrations of glycated albumin that are found in clinical specimensfrom human subjects, and do not depend on elevated glucoseconcentrations to be operative. Since the circulating half-life ofalbumin in humans is ≈17 days, there is prolonged exposure of vascularbeds to the glycated protein after it is formed. The use of agents thatblock the effects of glycated albumin to ameliorate vascular pathologieshas been explored in several scientific studies (Kidney International45:1673-1679, 1994; Journal of Clinical Investigation 95:2338-2345,1995; Diabetologia 39:270-274, 1996; Journal of Diabetic Complications12:28-33, 1998). Such agents may be monoclonal antibodies or othermolecules that react specifically with fructosyllysine residues that arepresent on glycated albumin but are not present on nonglycated albumin,and which are disclosed in U.S. Pat. No. 5,223,392 and U.S. Pat. No.5,518,720, incorporated by reference herein. Such therapies have beenshown, among other things, to prevent the structural and functionalchanges characteristic of renal and retinal microvascular disease.Anti-glycated albumin therapy therefore has been proposed as a treatmentmodality for vascular pathologies.

[0005] A novel approach to prevent pathologies related to the biologiceffects of glycated albumin would be to reduce the formation of theglycated protein and to lower its concentration in the circulation. Thiscould be accomplished by administering compounds that, by binding tospecific sites in albumin, can inhibit the attachment of glucose tophysiologically important lysine amino groups. The compounds wouldachieve this desired effect by obscuring the reactive lysine amino groupand/or causing a conformational shift in the tertiary structure of thealbumin molecule that renders the important glycatable siteinaccessible.

[0006] Identification of compounds which prevent glycation atphysiologically important sites is difficult to accomplish and has notbeen described in the art. In vitro glycation is distinctly differentfrom in vivo glycation. Excessive concentrations of glucose or reducingcarbohydrate are used, and the number of sites that undergo glycation issignificantly increased relative to sites which are subject to glycationin vivo. Physiologically important sites are only a small subset of thetotal number of sites and cannot be distinguished from the unimportantones after in vitro glycation according to methods described in the art.

[0007] Binding to albumin is a likely prerequisite for a compound toprevent glycation of albumin and many compounds of diverse structuralclasses have been shown to bind to albumin at various sites. Examplesinclude: vitamin C, vitamin E, vitamin B₆, diclofenac, acetylsalicylicacid, warfarin, bilirubin, iodobenzoic acids, diazepam, digitoxin,clofibrate, methotrexate, lithium, phenobarbital, cyclosporinbenzodiazepine, paracetamol, ibuprofen, suprofen, aminodarone,propanolol, griseofulvan, and others. But binding to albumin is notsufficient for antiglycation activity. Only a few compounds have beenreported to influence the condensation of carbohydrate with reactiveprotein amino groups in vitro, and none of them have been shown toaffect lysine amino groups that are physiologically important in vivo orto be of therapeutic benefit when administered in vivo with respect toglycation-related pathologies (Biochemical & Biophysical ResearchCommunications 165:991-996, 1988; Life Sciences 43:1725-1731, 1988;Diabete & Metabolisme 14:40-42, 1988; Biochemica et Biophysica Acta1120:201-204, 1992; Diabetes 41:167-173, 1992). Moreover, the conditionsemployed in such in vitro studies are irrelevant to in vivo conditionsas to degree of glycation, the reducing sugar and concentration used,and the concentration of compound tested. Binding to albumin andinhibition of in vitro glycation is not synonymous with prevention ofglycation at physiologically important sites.

[0008] It is a finding of the present invention that many compounds bindto albumin and inhibit glycation at unimportant sites but do not preventglycation at physiologically important sites.

[0009] It is another finding of the present invention that agents thatbind to fructosyllysine residues on albumin and, in so doing, preventpathobiologic effects of glycated albumin provide a useful tool forelucidating which albumin binding ligands are potentially important inpreventing the formation of glycated sites that are pathophysiologicallyimportant.

[0010] The present invention is directed toward discovery ofalbumin-binding compounds that block non-enzymatic glycation ofphysiologically important sites-which, when glycated, lead to vascularpathologies. The present invention is further directed to methods of useof these novel agents for the treatment of glycation-relatedpathologies, and novel methods of synthesis of these agents.

SUMMARY OF THE INVENTION

[0011] The present invention provides novel compounds that inhibit thenonenzymatic glycation of albumin at physiologically relevant sites.

[0012] The present invention also provides improved methods of synthesisof 2-phenylamino-phenylacetic acid derivatives.

[0013] The present invention further provides novel compositions forpreventing and treating glycation-induced pathologies.

[0014] These and other objects of the invention are achieved with thediscovery of compounds that are reactive with domain(s) in human albuminthat are susceptible to nonenzymatic glycation in vivo; that, by bindingto the sites in the structure of albumin, protect the protein fromattachment to glucose.

DETAILED DESCRIPTION

[0015] The present invention evolved from the finding thatglycation-associated pathologies can be ameliorated by ligand compoundsthat bind to specific glycated sites on albumin. A novel finding of thepresent invention is that these ligands can be used to identify othercompounds that prevent glycation of pathophysiologically important sitesin the albumin molecule that are selectively subject to glycation invivo and that, when glycated, cause deleterious biologic effects inrelevant tissues. The present invention further finds that thesecompounds can be identified by their ability to prevent, in theirbinding to albumin, the formation of fructosyllysine epitopes inglycated albumin that are recognized by monoclonal antibodies that aresite selective for fructosyl-lysine residues that are known to beinvolved in glycation-associated pathologies.

[0016] It is a finding of the present invention that compounds areidentified that, by binding to human albumin and protecting the proteinfrom nonenzymatic glycation at pathophysiologically important sites, aretherapeutically useful for the treatment of glycation-relatedpathologies.

[0017] Compounds of the present invention are capable of binding tosites in the primary structure of albumin which contain a lysine residuethat is a preferential site of nonenzymatic glycation in vivo and/orcause a conformational shift in the tertiary structure of the protein,rendering the glycatable site inaccessible to glucose attachment.

[0018] Compounds which are potentially useful are those which arecapable of binding albumin and include but are not limited to: vitaminC, vitamin E, vitamin B₆, diclofenac, acetylsalicylic acid, warfarin,bilirubin, iodobenzoic acids, diazepam, digitoxin, clofibrate,methotrexate, lithium, phenobarbital, cyclosporin benzodiazepine,paracetamol, ibuprofen, suprofen, aminodarone, propanolol, griseofulvan,and others.

[0019] A subset of therapeutically useful compounds can be identifiedwith the monoclonal antibody A717 which binds to fructosyl-lysineresidues on albumin and, in so doing, blocks the effects of glycatedalbumin on vascular pathology. Compounds which prevent the formation ofglycated sites recognized by A717 will be therapeutically useful.Compounds of the present invention possessing this activity are of thestructural formula:

[0020] wherein: X is hydrogen, sodium, lithium or potassium and R′₃,R′₄, R′₅, R′₆, R₂, R3, R₄, R₅, R₆ are the same or different and arehydrogen, chlorine, bromine, fluorine, iodine, methyl, ethyl, propyl,isopropyl, butyl, pentyl, butyloxy, pentyloxy, cyano, thio, methoxy,ethoxy, hydroxy, phosphate, sulfate, nitrate, or amino.

[0021] The compounds of the present invention can be tested and selectedfor low cyclo-oxygenase inhibitory activity and high anti-glycationactivity, since some compounds of this structural class have beenassociated with cyclo-oxygenase inhibitory activity. It is a finding ofthis invention that this property confers therapeutic advantage byachieving potent inhibition of albumin glycation and lessening untowardside-effects of cyclo-oxygenase inhibition when administered in vivo.

[0022] The compounds of the present invention are capable of preventingcellular and tissue damage that is evoked by glycated albumin that ispresent in the circulation. Since therapeutic concentrations of thecompounds of the present invention can inhibit the formation of glycatedalbumin with high IC₅₀ (the concentration giving 50% inhibition) ratiosof anti-glycation to cyclo-oxygenase inhibitory activities, the presentinvention provides a novel and improved method for the treatment ofglycation-related pathologies.

[0023] The compounds of this structural class(2-(phenylamino)phenylacetic acids) can be produced following themethods outlined in schemes 1-6. Scheme 1 describes the synthesis ofsubstituted diphenylamines and the subsequent condensation of theappropriate diphenylamines with refluxing chloracetyl chloride to yieldthe substituted 2-chloro-N-phenylacetanilides. Cyclization is achievedby heating at 160° C. in a melt with AlCl₃. Hydrolysis of thesubstituted N-aryloxindoles with NaOH in refluxing ethanolic solutionfollowed by acidification gives the 2-(phenylamino)phenylacetic acids.

[0024] Since alkyl migration and splitting of alkoxy groups can occurduring the cyclization reaction above, scheme 2 is beneficial in certaincases. In this scheme substituted diphenylamines are treated with oxalylchloride in benzene followed by cyclization of the N-phenyloxamic acidchloride with AlCl₃ in tetrachloroethane to yield N-arylisatins.Hydrolysis and acidification gives the corresponding phenylglyoxylicacids which are then reduced and acidified to produce the2-(phenylamino)phenylacetic acids.

[0025] Schemes 1 and 2 are general routes to 2-(phenylamino)phenylacetic acids, provided that both ortho positions of one phenyl ring ofthe diphenylamines are occupied in order to avoid the formation ofpositional isomers of the intermediate oxindoles and isatins. Schemes 3and 4 are synthetic routes that avoid the formation of isomers. Inscheme 3, potassium 2-iodophenyl acetate is reacted with substitutedanilines in the presence of potassium carbonate and activated copperpowder in hot N-methyl-2-pyrrolidone. Acidification and crystallizationyields the 2-(phenylamino) phenylacetic acids.

[0026] Scheme 4 involves the condensation of N,N-dimethyl-2-iodophenylacetamide and anilines in the presence ofanhydrous potassium carbonate, copper, and cuprous iodide in refluxingtoluene to give the substituted N,N-dimethyl-2-(phenylamino)phenylacetamides. Hydrolysis with KOH inrefluxing ethanol followed by acidification yields the2-(phenylamino)phenylacetic acids.

[0027] Hydroxylated 2-(phenylamio)phenylacetic acids are synthesizedfrom the appropriately substituted methoxy-2-(phenylamino)phenylaceticacids (prepared by scheme 2 and 4). In scheme 5, the methoxy-derivativesare treated with pyridine hydrochloride at 170° C. which gives thehydroxy substituted N-phenyloxindoles. Hydrolysis with NaOH in refluxingN-butanol completes the synthesis.

[0028] Hydroxylated compounds with additional methoxy groups areprepared by hydrogenation of he corresponding benzyloxy analogues withPd-C in tetrahydrofuran and 1,2-dichlorobenzene (scheme 6).

[0029] This invention provides an improved method of synthesis of2-(phenylamino)phenylacetic acids. Analogous to schemes 3 and 4, scheme7 produces the 2-(phenylamino)phenylacetic acid from a directcondensation of a phenylacetic acid with an aniline. In this scheme, thephenylacetic acid contains a reactive bromine, the amount of aniline isreduced, the reaction time is reduced, the potassium carbonate isreduced to prevent oxidation of the reaction products, theN-methylpyrrolidone is reduced to accelerate the bimolecular reaction,and reduced amounts of freshly prepared, activated copper is used. Thismethod improved yields of the desired product with little formation ofoxyindoles or oxidation products.

[0030] This invention also provides therapeutic compositions comprisingthe above-described compounds.

[0031] This invention further provides a method for treating diseasecomprising administering to the patient an effective amount of atherapeutic composition comprised of the above-described compound(s)capable of inhibiting albumin glycation and a pharmaceuticallyacceptable carrier therefor.

[0032] The present invention also comprises one or more compounds asdescribed above formulated into compositions together with one or morenon-toxic physiologically acceptable carriers, adjuvants or vehicleswhich are collectively referred to herein as carriers, for parenteralinjection for oral administration in solid or liquid form, for rectal ortopical administration, or the like. The compositions can beadministered to humans either orally, rectally, parenterally(intravenously, intramuscularly or subcutaneously), intracistemally,intravaginally, intraperitoneally, intravesically, locally (powders,ointments or drops), or as a buccal or nasal spray.

[0033] Compositions suitable for parenteral injection may comprisephysiologically acceptable sterile aqueous or nonaqueous solutions,dispersions, suspensions or emulsions and sterile powders forreconstitution into sterile injectable solutions or dispersions.Examples of suitable aqueous and nonaqueous carriers, diluents, solventsor vehicles include water, ethanol, polyols (propyleneglycol,polyethyleneglycol, glycerol, and the like), suitable mixtures thereof,vegetable oils (such as olive oil) and injectable organic esters such asethyl oleate. Proper fluidity can be maintained, for example, by the useof a coating such as lecithin, by the maintenance of the requiredparticle size in the case of dispersions and by the use of surfactants.

[0034] These compositions may also contain adjuvants such as preserving,wetting, emulsifying, and dispensing agents. Prevention of the action ofmicroorganisms can be ensured by various antibacterial and antifungalagents, for example, parabens, chlorobutanol, phenol, sorbic acid, andthe like. It may also be desirable to include isotonic agents, forexample sugars, sodium chloride and the like. Prolonged absorption ofthe injectable pharmaceutical form can be brought about by the use ofagents delaying absorption, for example, aluminum monostearate andgelatin.

[0035] Solid dosage forms for oral administration include capsules,tablets, pills, powders and granules. In such solid dosage forms, theactive compound is admixed with at least one inert customary,pharmaceutically acceptable, excipient (or carrier) such as sodiumcitrate or dicalcium phosphate or (a) fillers or extenders, as forexample, starches, lactose, sucrose, glucose, mannitol and silicic acid,(b) binders, as for example, carboxymethylcellulose, alginates, gelatin,polyvinylpyrrolidone, sucrose and acacia, (c) humectants, as forexample, glycerol, (d) disintegrating agents, as for example, agar-agar,calcium carbonate, potato or tapioca starch, alginic acid, certaincomplex silicates and sodium carbonate, (e) solution retarders, as forexample paraffin, (absorption accelerators, as for example, quaternaryammonium compounds, (g) wetting agents, as for example, cetyl alcoholand glycerol monostearate, (h) adsorbents, as for example, kaolin andbentonite, and (i) lubricants, as for example, talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate ormixtures thereof In the case of capsules, tablets and pills, the dosageforms may also comprise buffering agents.

[0036] Solid compositions of a similar type may also be employed asfillers in soft and hard-filled gelatin capsules using such excipientsas lactose or milk sugar as well as high molecular weightpolyethyleneglycols, and the like. Solid dosage forms such as tablets,capsules, pills and granules can be prepared with coatings and shells,such as enteric coatings and others well known in the art. They maycontain opacifying agents, and can also be of such composition that theyrelease the active compound or compounds in a certain part of theintestinal tract in a delayed manner. Examples of embedding compositionswhich can be used are polymeric substances and waxes.

[0037] The active compounds can also be in microencapsulated form, ifappropriate, with one or more of the above-mentioned excipients.

[0038] Liquid dosage forms for oral administration includepharmaceutically acceptable emulsions, solutions, suspensions, syrupsand elixirs. In addition to the active compounds, the liquid dosageforms may contain inert diluents commonly used in the art, such as wateror other solvents, solubilizing agents and emulsifiers, as for example,ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzylalcohol, benzyl benzoate, propyleneglycol, 1,3-butyleneglycol,dimethylformamide, oils, in particular, cottonseed oil, groundnut oil,corn germ oil, olive oil, castor oil and sesame oil, glycerol,tetrahydrofurfuryl alcohol, polyethyleneglycols and fatty acid esters ofsorbitan or mixtures of these substances, and the like.

[0039] Besides such inert diluents, the composition can also includeadjuvants, such as wetting agents, emulsifying and suspending agents,sweetening, flavoring and perfuming agents.

[0040] Suspensions, in addition to the active compounds, may containsuspending agents, as for example, ethoxylated isostearyl alcohols,polyethylene sorbitol and sorbitan esters, microcrystalline cellulose,aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixturesof these substances, and the like.

[0041] Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thepresent invention with suitable non-irritating excipients or carrierssuch as cocoa butter, polethyleneglycol or a suppository wax, which aresolid at ordinary temperatures but liquid at body temperature andtherefore, melt in the rectum or vaginal cavity and release the activecomponent.

[0042] Dosage forms for topical administration of a compound of thisinvention include ointments, powders, sprays and inhalants. The activecomponent is admixed under sterile conditions with a physiologicallyacceptable carrier and any preservatives, buffers or propellants as maybe required. Ophthalmic formulations, eye ointments, powders andsolutions are also contemplated as being within the scope of thisinvention.

[0043] Actual dosage levels of active ingredients in the compositions ofthe present invention may be varied so as to obtain an amount of activeingredient that is effective to obtain a desired therapeutic responsefor a particular composition and method of administration. The selecteddosage level therefore depends upon the desired therapeutic effect, onthe route of administration, on the desired duration of treatment andother factors.

[0044] The total daily dose of the compounds of this inventionadministered to a host in single or divided dose may be in amounts, forexample, of from about 1 nanomol to about 100 micromols per kilogram ofbody weight. Dosage unit compositions may contain such amounts or suchsubmultiples therefor as may be used to make up the daily dose. It willbe understood, however, that the specified dose level for any particularpatient will depend upon a variety of factors including the body weight,general health, sex, diet, time and route of administration, rates ofabsorption and excretion, combination with other drugs and the severityof the particular disease being treated. The dosage level may alsodepend on patient response as determined by measurement of theconcentration of glycated albumin in the circulation at suitableintervals after administration.

[0045] The above disclosure generally describes the present invention. Amore complete understanding can be obtained by reference to thefollowing specific examples which are provided herein for purposes ofillustration only, and are not intended to limit the scope of theinvention.

EXAMPLE 1

[0046] Identification of Compounds which Prevent the Glycation ofPathophysiologically Important Sites that are Subject to Glycation InVivo

[0047] Human albumin (10 mg/ml) was incubated for 4 days in bufferedsaline in the presence of 0 or 40 mM glucose, with or without the testcompound (1-1000 μM). After dialysis to remove free glucose andcompound, the amount of glycated albumin formed was determined byenzyme-linked immunosorbent assay using monoclonal antibodies known tospecifically react with Amadori-glucose adducts in glycated albumin andto be unreactive with the nonglycated protein (ATCC HB 9596; U.S. Pat.No. 5,223,392), incorporated by reference herein. In vivo administrationof this anti-glycated albumin antibody is also known to prevent theadverse biologic effects of glycated albumin. The performance of pairedincubations allowed comparison of the amount of glycated albumin formedunder control (0 mM glucose), stimulated (40 mM glucose) and inhibited(test compound plus glucose) conditions. From these data, compounds wereidentified which inhibit the formation of fructosyllysine epitopes inalbumin at lysine amino sites that are subject to glycation in vivo andthat are important for the biologic activity of glycated albumin. Arepresentative sample of test compounds from different structuralclasses is shown in Table 1. TABLE 1 Prevention of Albumin Glycation atPhysiologically Significant Sites IC₅₀ for Albumin Compound ClassGlycation (μM) 2[(2,6-dichlorophenyl) amino]- Heteroaryl  18 benzeneacetic acid acetic acid 1-methyl-5-[p-toluoyl] pyrrole- Heteroaryl  11.5× 10⁴ 2-acetic acid acetic acid 4-hydroxy-2-methyl-3-pyrid-[2- Enolicacid >10⁶ yl-carbamoyl] 1H-1,2 benzathiazine 1,1-dioxide2-[(2,6-dichloro-3-methyl- Anthranilic >10⁶ phenyl)-amino] benzoic acidacid α-methyl-4-[2-methyl) propyl)] Aryl propionic  7.8 × 10⁴ benzeneacetic acid acid 1-[p-chlorobenzoyl]-methoxy- Indole  2.7 × 10⁴2-methylindole-3-acetic acid acetic acid [Z]-5-fluoro-2-methyl-1 Indole 2.2 × 10⁴ [p-(methylsulfinyl) phenyl] acetic acid(methylene-1H-indene-3- acetic acid

[0048] Of these compounds, 2-[(2,6-dichlorophenyl)amino]phenylaceticacid showed the greatest ability to prevent glycation of thephysiologically important sites that are recognized by the site-specificantibodies reactive with fructosyllysine residues that are formed invivo as a result of nonenzymatic glycation.

EXAMPLE 2

[0049] Improved Synthesis of 2-(phenylamino)phenylacetic acids

[0050] One molar equivalent of bromophenylacetic acid or its cationicsalt, two equivalents of an appropriately substituted aniline, and twoequivalents of anhydrous potassium carbonate were mixed with 3 ml ofN-methyl pyrrolidine and to this mixture 7 mole percent of freshlyprepared activated copper was added. The reaction mixture was heated at115-120° C. for 4 hours. The resulting slightly grayish mixture wasfiltered hot over a bed of Celite and the Celite was washed with water(200 ml) and hexane (200 ml). The filtrate was transferred to aseparating funnel and extracted with hexane. The aqueous layer wascooled to 5° C. and neutralized with dilute hydrochloric acid (1:3)which also was kept at 5° C. The precipitated product was filtered,thoroughly washed with water, and dried under suction (30% yield basedon bromophenylacetic acid).

[0051] This protocol for successful synthesis of the desired compounddeparts from described procedures and provides improved methods forsynthesis of substituted anilines. Protocols described in the art employ1 equivalent of bromophenylacetic acid, 5 equivalents of the aniline, 4equivalents of anhydrous potassium carbonate, and 1 equivalent of copperpowder, that are mixed and refluxed in 30 ml methyl pyrrolidine for 20hours at 120° C., followed by treatment with hot water and thenchloroform to precipitate the potassium salt of the anilinophenylaceticacid. Application of the prior art methods yielded tarry reactionproducts that failed to form any precipitate, contained severalcompounds, and showed only a trace of the desired compound. The improvedmethod of synthesis a) reduces reaction time to 4 hours; b) maintainsreaction temperature between 115-120° C.; c) reduces the amount ofaniline to two equivalents per equivalent of bromophenylacetic acid; d)reduces the potassium carbonate to two equivalents (one to neutralizethe bromo acid and one for neutralizing the hydrobromic acid generatedin the reaction), since excess potassium carbonate caused base-catalyzedoxidation of the reaction products; e) reduces the amount of N-methylpyrrolidine 10 fold to accelerate the bimolecular reaction and enableisolation of the final product; and f) uses freshly activated copperinstead of copper powder, and reduces the amount to 7 mole percent ofthe bromophenylacetic acid used.

EXAMPLE 3

[0052] Synthesis of 2-[(phenyl)amino]phenylacetic acid

[0053] 2-bromophenylacetic acid (25 mmol) was added to a mixture of 50mmol of aniline, 50 mmol of anhydrous potassium carbonate, (7%) mmol ofactivated copper powder, and 3 ml of N-methylpyrrolidone at 120° C. Themixture was kept at 120° C. for 4 h with stirring. The resultingslightly grayish mixture was filtered while hot through a bed of Celiteand the Celite was washed with water (200 ml) and hexane (200 ml). Thefiltrate was transferred to a separating funnel and extracted withhexane and cooled to room temperature. The aqueous layer was removed,cooled to 5° C., and neutralized with dilute hydrochloric acid (1:3)which was also kept at 5° C. The precipitated2-[(phenyl)amino]phenylacetic acid was collected by filtration,thoroughly washed with water, and dried under suction.

EXAMPLE 4

[0054] Synthesis of 2-[(2-chlorophenyl)amino]phenylacetic acid

[0055] In the manner described in example 3, 2-bromophenylacetic acidwas condensed with 2-chloroaniline to yield2-[(2-chlorophenyl)amino]phenylacetic acid.

EXAMPLE 5

[0056] Synthesis of 2-[(3-chlorophenyl)amino]phenylacetic acid

[0057] In the manner described in example 3, 2-bromophenylacetic acidwas condensed with 3-chloroaniline to yield2-[(3-chlorophenyl)amino]phenylacetic acid.

[0058] NMR (DM 50-d₆: CH₂ singlet at 3.7 ppm; appropriate aromaticprotons with proper chemical shift; acid proton at 13 ppm

[0059] Elemental Analysis: C₁₄H₁₂ClNO₂ (acid); C₁₄H₁₁ClNO₂Na (salt)

[0060] TLC: Single spot

[0061] Melting point: 102-103° C.

EXAMPLE 6

[0062] Synthesis of 2-[(4-chlorophenyl)amino]phenylacetic acid

[0063] In the manner described in example 3,2-bromophenylacetic acid wascondensed with 4 chloroaniline to yield2-[(4-chlorophenyl)amino]phenylacetic acid.

EXAMPLE 7

[0064] Synthesis of 2-[(2,3-dichlorophenyl)amino]phenylacetic acid

[0065] In the manner described in example 3, 2-bromophenylacetic acidwas condensed with 2,3-dichloroaniline to yield2-[(2,3-dichlorophenyl)amino]phenylacetic acid.

EXAMPLE 8

[0066] Synthesis of 2-[(2,4-dichlorophenyl)amino]phenylacetic acid

[0067] In the manner described in example 3, 2-bromophenylacetic acidwas condensed with 2,4-dichloroaniline to yield2-[(2,4-dichlorophenyl)amino]phenylacetic acid.

EXAMPLE 9

[0068] Synthesis of 2-[(2,5-dichlorophenyl)amino]phenylacetic acid

[0069] In the manner described in example 3, 2-bromophenylacetic acidwas condensed with 2,5-dichloroaniline to yield2-[(2,5-dichlorophenyl)amino]phenylacetic acid.

EXAMPLE 10

[0070] Synthesis of 2-[(2,6-dichlorophenyl)amino]phenylacetic acid

[0071] In the manner described in example 3, 2-bromophenylacetic acidwas condensed with 2,6-dichloroaniline to yield2-[(2,6-dichlorophenyl)amino]phenylacetic acid.

EXAMPLE 11

[0072] Synthesis of 2-[(3,4-dichlorophenyl)amino]phenylacetic acid

[0073] In the manner described in example 3, 2-bromophenylacetic acidwas condensed with 3,4-dichloroaniline to yield2-[(3,4-dichlorophenyl)amino]phenylacetic acid.

EXAMPLE 12

[0074] Synthesis of 2-[(3,5-dichlorophenyl)amino]phenylacetic acid

[0075] In the manner described in example 3, 2-bromophenylacetic acidwas condensed with 3,5-dichloroaniline to yield2-[(3,5-dichlorophenyl)amino]phenylacetic acid.

EXAMPLE 13

[0076] Synthesis of 2-[(2,6-dimethylphenyl)amino]phenylacetic acid

[0077] In the manner described in example 3, 2-bromophenylacetic acidwas condensed with 2,6-dimethylaniline to yield2-[(2,6-dimethylphenyl]amino]phenylacetic acid.

EXAMPLE 14

[0078] Synthesis of 2-[(2,3-dimethylphenyl)amino]phenylacetic acid

[0079] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2,3-dimethylaniline to yield2-[(2,3-dimethylphenyl)amino]phenylacetic acid.

EXAMPLE 15

[0080] Synthesis of 2-[(2,4-dimethylphenyl)amino]phenylacetic acid

[0081] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2,4-dimethylaniline to yield2-[(2,4-dimethylphenyl)amino]phenylacetic acid.

EXAMPLE 16

[0082] Synthesis of 2-[(2,5-dimethylphenyl)amino]phenylacetic acid

[0083] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2,5-dimethylaniline to yield2-[(2,5-dimethylphenyl)amino]phenylacetic acid.

EXAMPLE 17

[0084] Synthesis of 2-[(3,4-dimethylphenyl)amino]phenylacetic acid

[0085] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3,4-dimethylaniline to yield2-[(3,4-dimethylphenyl)amino]phenylacetic acid.

EXAMPLE 18

[0086] Synthesis of 2-[(3,5-dimethylphenyl)amino]phenylacetic acid

[0087] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3,5-dimethylaniline to yield2-[(3,5-dimethylphenyl)amino]phenylacetic acid.

EXAMPLE 19

[0088] Synthesis of 2-[(2-methylphenyl)amino]phenylacetic acid

[0089] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with o-toluidine to yield2-[(2-methylphenyl)amino]phenylacetic acid.

EXAMPLE 20

[0090] Synthesis of 2-[(3-methylphenyl)amino]phenylacetic acid

[0091] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with m-toluidine to yield2-[(3-methylphenyl)amino]phenylacetic acid.

EXAMPLE 21

[0092] Synthesis of 2-[(4-methylphenyl)amino]phenylacetic acid

[0093] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with p-toluidine to yield2-[(4-methylphenyl)amino]phenylacetic acid.

EXAMPLE 22

[0094] Synthesis of 2-[(2,4,6-trichlorophenyl)amino]phenylacetic acid

[0095] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2,4,6-trichloroaniline to yield2-[(2,4,6-trichlorophenyl)amino]phenylacetic acid.

EXAMPLE 23

[0096] Synthesis of 2-[(2,6-dichloro-4-methoxyphenyl)amino]phenylaceticacid

[0097] Oxalyl chloride (0.5 mol) was added dropwise at 5° C. to asolution of 2,6-dichloro-4-methoxydiphenylamine (0.25 mol) in 375 ml ofbenzene. The mixture was stirred for 2 h at room temperature andevaporated. The residue was dissolved in 400 ml of benzene and thesolution was again evaporated to dryness to obtainN-(2,6-dichloro-4-methoxyphenyl)oxaniloyl chloride. This intermediatewas dissolved in 600 ml of tetrachloroethane. AlCl₃ (40 g) was addedslowly, and the mixture was stirred for 20 h at room temperature. Themixture was then poured over 200 ml of 2N HCl containing 800 g of ice.The organic phase was washed with water, 2N KHCO₃, again with water, andevaporated. Crystallization from ether yielded1-(2,6-dichloro-4-methoxyphenyl)isatin. This intermediate was dissolvedwin 215 ml of IN NaOH and 2100 ml of ethanol and was heated under refluxfor 10 min. The solution was cooled and evaporated. The residue wasdissolved in 2000 ml of water, washed with ether, and acidified with 2NHCl. The precipitate was extracted with ether. The organic extract gave,after washing with water, evaporation and crystallization for ether, theproduct 2-[2,6-dichloro4-methoxyphenyl)amino]glyoxylic acid. Thisintermediate (0.22 mol) was dissolved in 900 ml of 2-methoxyethanol.Hydrazine hydrate (1 mol) was added and the temperate of the mixture wasincreased to 60° C. NaOCH₃ (2.3 mol) was added slowly and the mixturewas slowly heated to 150° C., whereby methanol, water, hydrazine, andpart of the solvent evaporated. The mixture was kept at 150° C. for 1 h,collected, and poured over 8 kg of crushed ice. The aqueous phase wasextracted with 800 ml of ether and acidified with concentrated HCl at 0°C. The precipitated oil was extracted with ether. The ether extract waswashed with water and evaporated. The residue was crystallized fromether-petroleum ether to yielded2-[2,6-dichloro-4-methoxyphenyl)amino]phenylacetic acid.

EXAMPLE 24

[0098] Synthesis of 2-[(2,6-dichloro-4-hydroxyphenyl)amino]phenylaceticacid

[0099] The 2-[(2,6-dichloro-4-methoxyphenyl)amino]phenylacetic acid (0.1mol) from example 23 was added in portions to a melt of 200 g ofpyridine hydrochloride (1.75 mol) at 170° C. The mixture was heated at180° C. for 3 h and poured onto 2000 ml of ice water while hot. Theprecipitated product was filtered off, washed with water, and dissolvedin 1000 ml of ethyl acetate. The organic phase was washed with 200 ml ofIN HCl in water (2×100 ml) and evaporated to giveN-(2,6-dichloro-4-hydroxyphenyl)oxindole. To a solution of 8 g ofN-(2,6-dichloro-4-hydroxyphenyl)-oxindole in 200 ml of n-butanol wereadded 7 g of NaOH and 1 g of KOH, and the reaction mixture was refluxedfor 24 h and evaporated in vacuo. The residue was dissolved in 700 ml ofwater and the aqueous solution was extracted with ether (2×200 ml),cooled to 0° C., and acidified with concentrated HCl. The precipitatewas taken up in 300 ml of ether. The organic phase was washed with 30 mlof water, 0.5N NaHCO₃ (5×80 ml), and 80 ml of 2N KHCO₃ solution. Thecombined NaHCO₃ extracts were cooled to 0° C. and acidified with 2N HCl,and the precipitate was dissolved in 200 ml of ether. The organic layerwas washed with 30 ml of water and evaporated to yield2-[(2,6-dichloro-4-hydroxyphenyl)amino]phenylacetic acid.

EXAMPLE 25

[0100] Synthesis of 2-[(2,6-dichloro-3-methoxyphenyl)amino]phenylaceticacid

[0101] In the manner described in example 23,2,6-dichloro-3-methoxydiphenylamine was used to produce2-[(2,6-dichloro-3-methoxyphenyl)amino]phenylacetic acid.

EXAMPLE 26

[0102] Synthesis of 2-[(2,6-dichloro-3-hydroxyphenyl)amino]phenylaceticacid

[0103] In the manner described in example 24,2-[(2,6-dichloro-3-methoxyphenyl)amino]phenylacetic acid was convertedto 2-[(2,6-dichloro-3-hydroxyphenyl)amino]phenylacetic acid.

EXAMPLE 27

[0104] Synthesis of 2-[(2,6-dichloro-3-methylphenyl)amino]phenylaceticacid

[0105] A mixture of 2,6-dichloro-3-methyldiphenylamine (0.17 mol) andchloroacetyl chloride (0.5 mol) was refluxed for 16 h, cooled, andevaporated. The residue was dissolved in 500 ml of chloroform-ether(1:2). The organic phase was washed with 100 ml of 2N KHCO₃ and 100 mlof water and evaporated. The residue was recrystallized from MEOH togive 2-chloro-N-(2′,6′-dichloro-3′-methylphenyl)-N-phenylacetamide. Thisintermediate (0.1 mol) and 30 g of AlCl₃ were mixed, and the mixture washeated at 160° C. for 2 h (melting occurs at 100° C.). The molten masswas cooled and poured onto 300 g of crushed ice while the mixture wasstirred. The precipitated oil was dissolved in 300 ml of chloroform. Theorganic phase was washed with 50 ml of 2N KHO3 and 50 ml of water andevaporated. Recrystallization from MEOH gave1-(2,6-dichloro-3-methylphenyl) oxindole. A solution of 18.6 g of thisintermediate, 66 ml of 2N NaOH, and 66 ml of EtOH was refluxed for 4 h.The clear solution was cooled in an ice bath for 4 h. The precipitatedcrystals were filtered off and recrystallized from 80 ml of water toyield 2-[(2,6-dichloro-3-methylphenyl) amino]phenylacetic acid.

EXAMPLE 28

[0106] Synthesis of 2-[(2,6-dichlorophenyl)amino]5′-methoxyphenylaceticacid

[0107] In the manner described in example 23,4′-methoxy-2,6-dichlorodiphenylamine was used to yield2-[(2,6-dichlorophenyl)amino]5′-methoxyphenylacetic acid.

EXAMPLE 29

[0108] Synthesis of 2-[(2,6-dichlorophenyl)amino]5′-hydroxyphenylaceticacid.

[0109] In the manner described in example 24,2-[(2,6-dichlorophenyl)amino]5′-methoxyphenylacetic acid was convertedto 2-[(2,6-dichlorophenyl)amino]5′-hydroxyphenylacetic acid.

EXAMPLE 30

[0110] Synthesis of 2-[(2-methyl-3-chlorophenyl)amino]phenylacetic acid

[0111] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2-methyl-3-chloroaniline to yield2-[2-methyl-3-chlorophenyl)amino]phenylacetic acid.

EXAMPLE 31

[0112] Synthesis of 2-[(2,6-dichlorophenyl)amino]6′-bromophenylaceticacid

[0113] In the manner described in example 27,5-bromo-2′,6′-dichlorodiphenylamine is used to yield2-[(2,6-dichlorophenyl)amino]6′-bromophenylacetic acid.

EXAMPLE 32

[0114] Synthesis of 2-[(2-chloro-3-methylphenyl)amino]phenylacetic acid

[0115] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2-chloro-3-methylaniline to yield2-[(2-chloro-3-methylphenyl)amino]phenylacetic acid.

EXAMPLE 33

[0116] Synthesis of 2-[(2-chloro-6-fluorophenyl)amino]phenylacetic acid

[0117] In the manner described in example 27,2-chloro-6-fluorodiphenylamine is used to yield2-[(2-chloro-6-fluorophenyl)amino]phenylacetic acid.

EXAMPLE 34

[0118] Synthesis of2-[(2,3,5,6-tetramethylphenyl)amino]5′-chlorophenylacetic acid

[0119] In the manner described in example 23,4-chloro-2′,3′,5′,6′-tetramethyldiphenylamine is used to yield 2-[(2,3,5,6-tetramethylphenyl)amino]5′-chlorophenylacetic acid.

EXAMPLE 35

[0120] Synthesis of 2-[(2,6-diethylphenyl)amino]phenylacetic acid

[0121] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2,6-diethylaniline to yield2-[(2,6-diethylphenyl)amino]phenylacetic acid.

EXAMPLE 36

[0122] Synthesis of 2-[(2,4-difluorophenyl)amino]phenylacetic acid

[0123] In the manner described in example 3, 2-bromophenylacetic acidwas condensed with 2,4-difluoroaniline to yield2-[(2,4-difluorophenyl)amino]phenylacetic acid.

EXAMPLE 37

[0124] Synthesis of 2-[(2,6-difluorophenyl)amino]phenylacetic acid

[0125] In the manner described in example 3, 2-bromophenylacetic acidwas condensed with 2,6-difluoroaniline to yield2-[(2,6-difluorophenyl)amino]phenylacetic acid.

EXAMPLE 38

[0126] Synthesis of2-[(2,6-dichloro-3-methoxy4-benzyloxyphenyl)amino]phenylacetic acid

[0127] In the manner described in example 23,2,6-dichloro-3-methoxy-4-benzyloxydiphenylamine was used to yield2-[(2,6-dichloro-3-methoxy-4-benzyloxyphenyl)amino]phenylacetic acid.

EXAMPLE 39

[0128] Synthesis of2-[(2,6-dichloro-3-methoxy-4-hydroxyphenyl)amino]phenylacetic acid

[0129] The2-[(2,6-dichloro-3-methoxy-4-benzyloxyphenyl)amino]phenylacetic acid (10g) from example ,8 was hydrogenated with Pd-C (I g, 5%) intetrahydrofuran (100 ml) and 1,2-dichlorobenzene (10 ml) at normalpressure for 25 min at room temperature. The catalyst was removed byfiltration, and the filtrate was evaporated to leave the final product2-[(2,6-dichloro-3-methoxy-4-hydroxyphenyl) amino]phenylacetic acid.

EXAMPLE 40

[0130] Synthesis of 2-[(2,6-dichlorophenyl)amino]6′-methoxyphenylaceticacid

[0131] In the manner described in example 23,5-methoxy-2′,6′-dichlorodiphenylamine was used to yield2-[(2,6-dichlorophenyl)amino]6′-methoxyphenylacetic acid.

EXAMPLE 41

[0132] Synthesis of 2-[(2,6-dichlorophenyl)amino]6′-hydroxyphenylaceticacid

[0133] In the manner described in example 24,2-[(2,6-dichlorophenyl)amino]6′-methoxyphenylacetic acid was convertedto 2-[(2,6-dichlorophenyl)amino]6′-hydroxyphenylacetic acid.

EXAMPLE 42

[0134] Synthesis of2-[(2,6-dichloro-3-benzyloxy-4-methoxyphenyl)amino]phenylacetic acid

[0135] In the manner described in example 23,2,6-dichloro-3-benzyloxy-4-methoxydiphenylamine was used to yield2-[(2,6-dichloro-3-benzyloxy-4-methoxyphenyl)amino]phenylacetic acid.

EXAMPLE 43

[0136] Synthesis of2-[(2,6-dichloro-3-hydroxy-4-methoxyphenyl)amino]phenylacetic acid

[0137] In the manner described in example 39,2-[(2,6-dichloro-3-benzyloxy-4-methoxyphenyl)amino] phenylacetic acidwas converted to 2-[(2,6-dichloro-3-hydroxy-4-methoxyphenyl)amino]phenylacetic acid.

EXAMPLE 44

[0138] Synthesis of2-[(2,6-dichloro-4-methoxyphenyl)amino]5′-methoxyphenylacetic acid

[0139] In the manner described in example 23,4-methoxy-2′,6′-dichloro-4′-methoxydiphenylamine is used to yield2-[(2,6-dichloro-4-methoxyphenyl)amino]5′-methoxyphenylacetic acid.

EXAMPLE 45

[0140] Synthesis of2-[(2,6-dichloro-4-hydroxyphenyl)amino]5′-hydroxyphenylacetic acid

[0141] In the manner described in example 24,2-[(2,6-dichloro-4-methoxyphenyl)amino]5′-methoxyphenylacetic acid wasconverted to2-[(2,6-dichloro-4-hydroxyphenyl)amino]5′-hydroxyphenylacetic acid.

EXAMPLE 46

[0142] Synthesis of 2-[(2-methoxyphenyl)amino]phenylacetic acid

[0143] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with o-anisidine to yield2-[(2-methoxyphenyl)amino]phenylacetic acid.

EXAMPLE 47

[0144] Synthesis of 2-[(3-methoxyphenyl)amino]phenylacetic acid

[0145] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with m-anisidine to yield2-[(3-methoxyphenyl)amino]phenylacetic acid.

EXAMPLE 48

[0146] Synthesis of 2-[(4-methoxyphenyl)amino]phenylacetic acid

[0147] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with p-anisidine to yield2-[(4-methoxyphenyl)amino]phenylacetic acid.

EXAMPLE 49

[0148] Synthesis of 2-[(2-hydroxyphenyl)amino]phenylacetic acid

[0149] In the manner described in example 24,2-[(2-methoxyphenyl)amino]phenylacetic acid is converted to2-[(2-hydroxyphenyl)amino]phenylacetic acid.

EXAMPLE 50

[0150] Synthesis of 2-[(3-hydroxyphenyl)amino]phenylacetic acid

[0151] In the manner described in example 24,2-[(3′-methoxyphenyl)amino]phenylacetic acid is converted to2-[(3-hydroxyphenyl)amino]phenylacetic acid.

EXAMPLE 51

[0152] Synthesis of 2-[(4-hydroxyphenyl)amino]phenylacetic acid

[0153] In the manner described in example 24,2-[(4-methoxyphenyl)amino]phenylacetic acid is converted to2-[(4-hydroxyphenyl)amino]phenylacetic acid.

EXAMPLE 52

[0154] Synthesis of 2-[(3-chloro-4-methoxyphenyl)amino]phenylacetic acid

[0155] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3-chloro-4-methoxyaniline to yield2-[(=—chloro-4-methoxyphenyl)amino]phenylacetic acid.

EXAMPLE 53

[0156] Synthesis of 2-[(3-chloro-4-hydroxyphenyl)amino]phenylacetic acid

[0157] In the manner described in example 24,2-[(3-chloro-4-methoxyphenyl)amino]phenylacetic acid is converted to2-[(3-chloro-4-hydroxyphenyl)amino]phenylacetic acid.

EXAMPLE 54

[0158] Synthesis of 2-[(2-methoxy-5-chlorophenyl)amino]phenylacetic acid

[0159] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2-methoxy-5-chloroaniline to yield2-[(2-methoxy-5-chlorophenyl)amino]phenylacetic acid.

EXAMPLE 55

[0160] Synthesis of 2-[(2-hydroxy-5-chlorophenyl)amino]phenylacetic acid

[0161] In the manner described in example 24,2-[(2-methoxy-5-chlorophenyl)amino]phenylacetic acid is converted to2-[(2-hydroxy-5-chlorophenyl)amino]phenylacetic acid.

EXAMPLE 56

[0162] Synthesis of 2-[(3-methoxy-6-chlorophenyl)amino]phenylacetic acid

[0163] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3-methoxy-6-chloroaniline to yield2-[(3-methoxy-6-chlorophenyl)amino]phenylacetic acid.

EXAMPLE 57

[0164] Synthesis of 2-[(3-hydroxy-6-chlorophenyl)amino]phenylacetic acid

[0165] In the manner described in example 24,2-[(3-methoxy-6-chlorophenyl)amino]phenylacetic acid is converted to2-[(3-hydroxy-6-chlorophenyl)amino]phenylacetic acid.

EXAMPLE 58

[0166] Synthesis of 2-[(2-methoxy-3-fluorophenyl)amino]phenylacetic acid

[0167] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2-methoxy-3-fluoroaniline to yield2-[(2-methoxy-3-fluorophenyl)amino]phenylacetic acid.

EXAMPLE 59

[0168] Synthesis of 2-[(2-hydroxy-3-fluorophenyl)amino]phenylacetic acid

[0169] In the manner described in example 24,2-[(2-methoxy-3-fluorophenyl)amino]phenylacetic acid is converted to2-[(2-hydroxy-3-fluorophenyl)amino]phenylacetic acid.

EXAMPLE 60

[0170] Synthesis of 2-[(3-fluoro-4-methoxyphenyl)amino]phenylacetic acid

[0171] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3-fluoro-4-methoxyaniline to yield2-[(3-fluoro-4-methoxyphenyl)amino]phenylacetic acid.

EXAMPLE 61

[0172] Synthesis of 2-[(3-fluoro-4-hydroxyphenyl)amino]phenylacetic acid

[0173] In the manner described in example 24,2-[(3-fluoro-4-methoxyphenyl)amino]phenylacetic acid is converted to2-[(3-fluoro-4-hydroxyphenyl)amino]phenylacetic acid.

EXAMPLE 62

[0174] Synthesis of 2-[(2-methoxy-4-nitrophenyl)amino]phenylacetic acid

[0175] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2-methoxy-4-nitroaniline to yield2-[(2-methoxy4-nitrophenyl)amino]phenylacetic acid.

EXAMPLE 63

[0176] Synthesis of 2-[(2-hydroxy-4-nitrophenyl)amino]phenylacetic acid

[0177] In the manner described in example 24,2-[(2-methoxy-4-nitrophenyl)amino]phenylacetic acid is converted to2-[(2-hydroxy-4-nitrophenyl)amino]phenylacetic acid.

EXAMPLE 64

[0178] Synthesis of 2-[(2-methoxy-5-nitrophenyl)amino]phenylacetic acid

[0179] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2-methoxy-5-nitroaniline to yield2-[(2-methoxy-5-nitrophenyl)amino]phenylacetic acid.

EXAMPLE 65

[0180] Synthesis of 2-[(2-hydroxy-5-nitrophenyl)amino]phenylacetic acid

[0181] In the manner described in example 24,2-[(2-methoxy-5-nitrophenyl)amino]phenylacetic acid is converted to2-[(2-hydroxy-5-nitrophenyl)amino]phenylacetic acid.

EXAMPLE 66

[0182] Synthesis of 2-[(2-nitro-4-methoxyphenyl)amino]phenylacetic acid

[0183] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2-nitro-4-methoxyaniline to yield2-[(2-nitro-4-methoxyphenyl)amino]phenylacetic acid.

EXAMPLE 67

[0184] Synthesis of 2-[(2-nitro-4-hydroxyphenyl)amino]phenylacetic acid

[0185] In the manner described in example 24,2-[(2-nitro-4-methoxyphenyl)amino]phenylacetic acid is converted to2-[(2-nitro-4-hydroxyphenyl)amino]phenylacetic acid

EXAMPLE 68

[0186] Synthesis of 2-[(2-nitro-4-ethoxyphenyl)amino]phenylacetic acid

[0187] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2-nitro-4-ethoxyaniline to yield2-[(2-nitro-4-ethoxyphenyl)amino]phenylacetic acid.

EXAMPLE 69

[0188] Synthesis of2-[(3-methoxy-5-(trifluoromethyl)phenyl)amino]phenylacetic acid

[0189] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3-methoxy-5-(trifluoromethyl)aniline to yield2-[(3-methoxy-5-(trifluoromethyl)phenyl)amino]phenylacetic acid.

EXAMPLE 70

[0190] Synthesis of2-[(3-hydroxy-5-(trifluoromethyl)phenyl)amino]phenylacetic acid

[0191] In the manner described in example 24,2-[(3-methoxy-5-(trifluoromethyl)phenyl)amino] phenylacetic acid isconverted to 2-[(3-hydroxy-5-(trifluoromethyl)phenyl)amino]phenylaceticacid.

EXAMPLE 71

[0192] Synthesis of 2-[(2-ethylphenyl)amino]phenylacetic acid

[0193] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2-ethylaniline to yield2-[(2-ethylphenyl)amino]phenylacetic acid.

EXAMPLE 72

[0194] Synthesis of 2-[(3-ethylphenyl)amino]phenylacetic acid

[0195] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3-ethylaniline to yield2-[(3-ethylphenyl)amino]phenylacetic acid.

EXAMPLE 73

[0196] Synthesis of 2-[(4-ethylphenyl)amino]phenylacetic acid

[0197] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 4-ethylaniline to yield2-[(4-ethylphenyl)amino]phenylacetic acid.

EXAMPLE 74

[0198] Synthesis of 2-[(2-bromophenyl)amino]phenylacetic acid

[0199] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2-bromoaniline to yield2-[(2-bromophenyl)amino]phenylacetic acid.

EXAMPLE 75

[0200] Synthesis of 2-[(3-bromophenyl)amino]phenylacetic acid

[0201] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3-bromoaniline to yield2-[(3-bromophenyl)amino]phenylacetic acid.

EXAMPLE 76

[0202] Synthesis of 2-[(4-bromophenyl)amino]phenylacetic acid

[0203] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 4-bromoaniline to yield2-[(4-bromophenyl)amino]phenylacetic acid.

EXAMPLE 77

[0204] Synthesis of 2-[(2-fluorophenyl)amino]phenylacetic acid

[0205] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2-fluoroaniline to yield2-[(2-fluorophenyl)amino]phenylacetic acid.

EXAMPLE 78

[0206] Synthesis of 2-[(3-fluorophenyl)amino]phenylacetic acid

[0207] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3-fluoroaniline to yield2-[(3-fluorophenyl)amino]phenylacetic acid.

EXAMPLE 79

[0208] Synthesis of 2-[(4-fluorophenyl)amino]phenylacetic acid

[0209] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 4-fluoroaniline to yield2-[(4-fluorophenyl)amino]phenylacetic acid.

EXAMPLE 80

[0210] Synthesis of 2-[(2-iodophenyl amino]phenylacetic acid

[0211] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2-iodoaniline to yield2-[(2-iodophenyl)amino]phenylacetic acid.

EXAMPLE 81

[0212] Synthesis of 2-[(3-iodophenyl)amino]phenylacetic acid

[0213] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3-iodoaniline to yield2-[(3-iodophenyl)amino]phenylacetic acid.

EXAMPLE 82

[0214] Synthesis of 2-[(4-iodophenyl)amino]phenylacetic acid

[0215] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 4-iodoaniline to yield2-[(4-iodophenyl)amino]phenylacetic acid.

EXAMPLE 83

[0216] Synthesis of 2-[(2-nitrophenyl)amino]phenylacetic acid

[0217] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2-nitroaniline to yield2-[(2-nitrophenyl)amino]phenylacetic acid.

EXAMPLE 84

[0218] Synthesis of 2-[(3-nitrophenyl)amino]phenylacetic acid

[0219] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3-nitroaniline to yield2-[(3-nitrophenyl)amino]phenylacetic acid.

EXAMPLE 85

[0220] Synthesis of 2-[(4-nitrophenyl)amino]phenylacetic acid

[0221] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 4-nitroaniline to yield2-[(4-nitrophenyl)amino]phenylacetic acid.

EXAMPLE 86

[0222] Synthesis of 2-[(3,4-difluorophenyl)amino]phenylacetic acid

[0223] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3,4-difluoroaniline to yield2-[(3,4-difluorophenyl)amino]phenylacetic acid.

EXAMPLE 87

[0224] Synthesis of 2-[(3,5-difluorophenyl)amino]phenylacetic acid

[0225] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3,5-difluoroaniline to yield2-[(3,5-difluorophenyl)amino]phenylacetic acid.

EXAMPLE 88

[0226] Synthesis of 2-[(2,5-difluorophenyl)amino]phenylacetic acid

[0227] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2,5-difluoroaniline to yield2-[(2,5-difluorophenyl)amino]phenylacetic acid.

EXAMPLE 89

[0228] Synthesis of 2-[(2,3-difluorophenyl)amino]phenylacetic acid

[0229] In the manner described in example 3 2-bromophenylacetic acid iscondensed with 2,3-difluoroaniline to yield2-[(2,3-difluorophenyl)amino]phenylacetic acid.

EXAMPLE 90

[0230] Synthesis of 2-[(2,4-dibromophenyl)amino]phenylacetic acid

[0231] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2,4-dibromoaniline to yield2-[(2,4-dibromophenyl)amino]phenylacetic acid.

EXAMPLE 91

[0232] Synthesis of 2-[(2,5-dibromophenyl)amino]phenylacetic acid

[0233] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2,5-dibromoaniline to yield2-[(2,5-dibromophenyl)amino]phenylacetic acid.

EXAMPLE 92

[0234] Synthesis of 2-[(2,6-dibromophenyl)amino]phenylacetic acid

[0235] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2,6-dibromoaniline to yield2-[(2,6-dibromophenyl)amino]phenylacetic acid.

EXAMPLE 93

[0236] Synthesis of 2-[(3-chloro-4-fluorophenyl)amino]phenylacetic acid

[0237] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3-chloro-4-fluoroaniline to yield2-[(3-chloro-4-fluorophenyl)amino]phenylacetic acid.

EXAMPLE 94

[0238] Synthesis of 2-[(2-fluoro-4-chlorophenyl)amino]phenylacetic acid

[0239] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2-fluoro-4-chloroaniline to yield2-[(2-fluoro-4-chlorophenyl)amino]phenylacetic acid.

EXAMPLE 95

[0240] Synthesis of 2-[(3-nitro-4-chlorophenyl)amino]phenylacetic acid

[0241] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3-nitro-4-chloroaniline to yield2-[(3-nitro-4-chlorophenyl)amino]phenylacetic acid.

EXAMPLE 96

[0242] Synthesis of 2-[(2-fluoro-5-nitrophenyl)amino]phenylacetic acid

[0243] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2-fluoro-5-nitroaniline to yield2-[(2-fluoro-5-nitrophenyl)amino]phenylacetic acid.

EXAMPLE 97

[0244] Synthesis of 2-[(3-nitro-4-fluorophenyl)amino]phenylacetic acid

[0245] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3-nitro-4-fluoroaniline to yield2-[(3-nitro-4-fluorophenyl)amino]phenylacetic acid.

EXAMPLE 98

[0246] Synthesis of 2-[(2-fluoro-4-iodophenyl)amino]phenylacetic acid

[0247] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2-fluoro-4-iodoaniline to yield2-[(2-fluoro-4-iodophenyl)amino]phenylacetic acid.

EXAMPLE 99

[0248] Synthesis of 2-[(3,5-dinitrophenyl)amino]phenylacetic acid

[0249] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3,5-dinitroaniline to yield2-[(3,5-dinitrophenyl)amino]phenylacetic acid.

EXAMPLE 100

[0250] Synthesis of 2-[(2-fluoro-4-bromophenyl)amino]phenylacetic acid

[0251] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2-fluoro-4-bromoaniline to yield2-[(2-fluoro-4-bromophenyl)amino]phenylacetic acid.

EXAMPLE 101

[0252] Synthesis of 2-[(2,3,4-trifluorophenyl)amino]phenylacetic acid

[0253] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2,3,4-trifluoroaniline to yield2-[(2,3,4-trifluorophenyl)amino]phenylacetic acid.

EXAMPLE 102

[0254] Synthesis of 2-[(3,4,5-trichlorophenyl)amino]phenylacetic acid

[0255] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3,4,5-trichloroaniline to yield2-[(3,4,5-trichlorophenyl)amino]phenylacetic acid.

EXAMPLE 103

[0256] Synthesis of 2-[(2,4,5-triflourophenyl)amino]phenylacetic acid

[0257] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2,4,5-triflouroaniline to yield2-[(2,4,5-triflourophenyl)amino]phenylacetic acid.

EXAMPLE 104

[0258] Synthesis of 2-[(2,3,4,6-tetrafluorophenyl)amino]phenylaceticacid

[0259] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 2,3,4,6-tetrafluoroaniline to yield2-[(2,3,4,6-tetrafluorophenyl)amino]phenylacetic acid.

EXAMPLE 105

[0260] Synthesis of 2-[(3-methyl-4-bromophenyl)amino]phenylacetic acid

[0261] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3-methyl-4-bromoaniline to yield2-[(3-methyl-4-bromophenyl)amino]phenylacetic acid.

EXAMPLE 106

[0262] Synthesis of 2-[(3-bromo-4-methylphenyl)amino]phenylacetic acid

[0263] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3-bromo-4-methylaniline to yield2-[(3-bromo-4-methylphenyl)amino]phenylacetic acid.

EXAMPLE 107

[0264] Synthesis of 2-[(3-fluoro-4-methylphenyl)amino]phenylacetic acid

[0265] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3-fluoro-4-methylaniline to yield2-[(3-fluoro-4-methylphenyl)amino]phenylacetic acid.

EXAMPLE 108

[0266] Synthesis of 2-[(3-methylmercaptophenyl)amino]phenylacetic acid

[0267] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3-methylmercaptoaniline to yield2-[(3-methylmercaptophenyl)amino]phenylacetic acid.

EXAMPLE 109

[0268] Synthesis of 2-[(4-methylmercaptophenyl)amino]phenylacetic acid

[0269] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 4-methylmercaptoaniline to yield2-[(4-methylmercaptophenyl)amino]phenylacetic acid.

EXAMPLE 110

[0270] Synthesis of 2-[(3-nitro-4-methylphenyl)amino]phenylacetic acid

[0271] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3-nitro-4-methylaniline to yield2-[(3-nitro-4-methylphenyl)amino]phenylacetic acid.

EXAMPLE 111

[0272] Synthesis of 2-[(3,5-methoxyphenyl)amino]phenylacetic acid

[0273] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3,5-methoxyaniline to yield2-[(3,5-methoxyphenyl)amino]phenylacetic acid.

EXAMPLE 112

[0274] Synthesis of 2-[(3,5-hydroxyphenyl)amino]phenylacetic acid

[0275] In the -manner described in example 24,2-[(3,5-methoxyphenyl)amino]phenylacetic acid is converted to2-[(3,5-hydroxyphenyl)amino]phenylacetic acid

EXAMPLE 113

[0276] Synthesis of 2-[(4-propylphenyl)amino]phenylacetic acid

[0277] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 4-propylaniline to yield2-[(4-propylphenyl)amino]phenylacetic acid.

EXAMPLE 114

[0278] Synthesis of 2-[(4-isopropylphenyl)amino]phenylacetic acid

[0279] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 4-isopropylaniline to yield2-[(4-isopropylphenyl)amino]phenylacetic acid.

EXAMPLE 115

[0280] Synthesis of 2-[(3,4,5-trimethoxyphenyl)amino]phenylacetic acid

[0281] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 3,4,5-trimethoxyaniline to yield2-[(3,4,5-trimethoxyphenyl)amino]phenylacetic acid.

EXAMPLE 116

[0282] Synthesis of 2-[(3,4,5-trihydroxyphenyl)amino]phenylacetic acid

[0283] In the manner described in example 24,2-[(3,4,5-trimethoxyphenyl)amino]phenylacetic acid is converted to2-[(3,4,5-trihydroxyphenyl)amino]phenylacetic acid.

EXAMPLE 117

[0284] Synthesis of 2-[(4-butylphenyl)amino]phenylacetic acid

[0285] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 4-butylaniline to yield2-[(4-butylphenyl)amino]phenylacetic acid.

EXAMPLE 118

[0286] Synthesis of 2-[(4-butoxyphenyl)amino]phenylacetic acid

[0287] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 4-butoxyaniline to yield2-[(4-butoxyphenyl)amino]phenylacetic acid.

EXAMPLE 119

[0288] Synthesis of 2-[(4-pentylphenyl)amino]phenylacetic acid

[0289] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 4-pentylaniline to yield2-[(4-pentylphenyl)amino]phenylacetic acid.

EXAMPLE 120

[0290] Synthesis of 2-[(4-pentyloxyphenyl)amino]phenylacetic acid

[0291] In the manner described in example 3, 2-bromophenylacetic acid iscondensed with 4-pentyloxyaniline to yield2-[(4-pentyloxyphenyl)amino]phenylacetic acid.

EXAMPLE 121

[0292] Anti-Glycation Activity of 2-(phenylamino)phenylacetic acids

[0293] Human albumin (10 mg/ml) was incubated for 48-144 hours at 37° C.in the presence of 0-50 mM glucose in buffered saline, without or withthe test compound in varying concentrations (1-1000 μM). After dialysisto remove free glucose and compound, the amount of glycated albuminformed was determined by enzyme-linked immunosorbent assay using themonoclonal antibodies known to specifically react with Amadori-glucoseadducts in glycated albumin and to be unreactive with the nonglycatedprotein. The presence of glucose in the incubations promotes thenonenzymatic glycation of albumin, and the performance of pairedincubations allows comparison of the amount of glycated albumin formedunder control (0 glucose), stimulated (25-50 mM glucose), and inhibited(compound plus glucose) conditions. From these data, the IC₅₀(concentration required for 50% inhibition) for glycation inhibition wascalculated. The IC₅₀ (μM) for inhibition of glycation by arepresentative sample of 2-(phenylamino)phenylacetic acid compounds isshown in Table 2. TABLE 2 Prevention of Albumin Glycation by2-(phenylamino) phenylacetic acids Example Glycation Inhibition # IC₅₀(μM) 3 155 4 10 5 50 6 11 8 63 10 6.3 11 2.5 13 112

EXAMPLE 122

[0294] Cyclo-oxygenase Inhibitory Activity of2-(phenylamino)phenylacetic acids

[0295] Cellular homogenates of bovine seminal vesicles were incubatedwith arachidonic acid and the production of prostaglandins wasmonitored. The effect of test compound on the conversion of -arachidonicacid to prostaglandin E₂ was measured after incubation for 1-30 minutesin buffer containing the cofactors hematin and phenol. Compounds weretested in varying concentration (1-1000 μM) and the data used tocalculate the IC₅₀ for cyclo-oxygenase inhibition. Thecyclo-oxygenase-inhibitory activity of representative2-(phenylamino)phenylacetic acids is presented in Table 3. TABLE 3 COXInhibitory Activity of 2-(phenylamino) phenylacetic acids. Example COXInhibition # IC50 (μM) 3 288 4 42 5 60 6 98 8 71 10 1 11 141 13 6

EXAMPLE 123

[0296] Relative Anti-Glycation versus Cyclo-oxygenase InhibitoryActivity

[0297] It was determined from the ratio of 6.3 of the IC₅₀ of glycationto cyclo-oxygenase inhibitory activities of2-(2,6-dichlorophenylamino)phenylacetic acid that an anti-glycation:cyclo-oxygenase inhibition IC₅₀ ratio less than 2 would provide afavorable therapeutic profile in which therapeutic concentrationsachieving significant anti-glycation activity would have insignificantcyclo-oxygenase inhibitory activity. The anti-glycation: cyclo-oxygenaseinhibition ratio of representative 2-(phenylamino)phenylacetic acids isshown in Table 4. TABLE 4 Example Antiglycation IC₅₀ versus # COXInhibition IC₅₀ 3 0.54 4 0.24 5 0.80 6 0.11 8 0.90 10 6.3 11 0.02 1319.0

EXAMPLE 124

[0298] Prevention of Albumin Glycation In Vivo by A-Administration of2-(phenylamino) phenylacetic acids.

[0299] Mice were administered 3 to 10 mg/kg of2-(2-chlorophenylamino)phenylacetic acid,2-(3-chlorophenylamino)phenylacetic acid, or 2-(2,6dichlorophenylamino)phenylacetic acid, given in divided doses byintraperitoneal injection, for five days. Blood was taken before theinitiation of treatment and four hours after the last dosing at theconclusion of the treatment. After centrifugation to separate plasmafrom red cells, the plasma concentration of glycated albumin wasdetermined by enzyme-linked immunosorbent assay using the monoclonalantibodies described in Example 1. As shown below, all of the2-(phenylamino)phenylacetic acids tested produced a dose-relatedreduction in plasma glycated albumin concentrations. Glycated Albumin(μg/ml) Percent Dose Pre-treatment Post-treatment change2-(3-chlorophenylamino) phenylacetic acid  5 mg/kg/day 854 697 −18.4 10mg/kg/day 647 352 −45.6 2-(2-chlorophenylamino) phenylacetic acid  5mg/kg/day 741 500 −32.5 10 mg/kg/day 815 426 −47.72-(2,6-dichlorophenylamino) phenylacetic acid  3 mg/kg/day 841 760 −9.6 6 mg/kg/day 946 690 −27.1

EXAMPLE 125

[0300] Lowering Glycated Albumin with 2-(phenylamino)phenyl acetic acidcompounds ameliorates glycation-related pathology

[0301] Mice were treated for 8 weeks with 6 mg/kg/day of2-(2,6-dichlorophenylamino)phenylacaetic acid in divided doses. Urinesamples were obtained at the initiation and termination of treatment forexamination of protein excretion, which measures vascular dysfunctionthat is associated with glycation-related pathology As shown below, thereduction in glycated albumin was associated with a reduction in urineprotein excretion. Urine protein μg/24 hrs Initiation 7.6 Termination2.7

EXAMPLE 126

[0302] Therapeutic Composition/Treatment

[0303] A. Tablet

[0304] A typical tablet contains 2-[(2-chlorophenyl)amino]phenylaceticacid (100 mg), pregelatirlized starch USP (82 mg), microcrystallinecellulose (82 mg), and magnesium stearate (1 mg). In like manners, forexample, 2-[(3-chlorophenyl)amino]phenylacetic acid can be formulated inplace of 2-[(2-chlorophenyl)amino]phenyl acetic acid.

[0305] B. Liquid

[0306] A typical liquid contains 2-[(2-chlorophenyl)amino]phenylaceticacid (50 mg), sodium phosphate dibasic (50 mg), ethyl alcohol (0.5 ml),water (5 ml) and sweetening and/or flavoring agents. Similarly, otherformulations can be made by substituting, for example,2-[(3-chlorophenyl) amino]phenylacetic acid for2-[(2-chlorophenyl)amino]phenylacetic acid.

[0307] C. Injection

[0308] A typical injectable formulation contains2-[(2-chlorophenyl)amino]phenylacetic acid (25 mg) sodium phosphatedibasic (11.4 mg), benzyl alcohol (0.01 ml) and water for injection (1ml). Similarly, this formulation can be prepared employing, for example,2-[(3 chlorophenyl)amino] phenylacetic acid in place of 2-[(2chlorophenyl)amino]phenylacetic acid.

[0309] D. Suppositories

[0310] Typical suppository formulations can contain 2-[(2chlorophenyl)amino]phenylacetic acid (50 mg) butylated hydroxyanisol(0.1-1.0 mg), disodium calcium edetate (0.25-0.50 mg), andpolyethyleneglycol (775-1600 mg). Other suppository formulations can bemade by substituting, for example, 2-[(3-chlorophenyl)amino]phenylaceticacid for 2-[(2-chlorophenyl)amino]phenylacetic acid and by substituting,for example, butylated hydroxytoluene (0.04-0.08 mg) for the disodiumcalcium edetate and a hydrogenated vegetable oil (678-1400 mg), such asSuppocire L, Wecobee FS, Wecobee M, Witepsols, and the like, for thepolyethylene glycol.

I claim:
 1. A compound that inhibits the formation of glucose adducts onalbumin.
 2. A compound of claim 1 that inhibits the formation of glucoseadducts on one or more residues of albumin as defined by a monoclonalantibody produced by cell line ATCC HB9596.
 3. A compound selected fromthose having the following structure:

wherein: R′₃, R′₄, R′₅, R′₆, R₂, R₃, R₄, R₅, and R₆, are the same ordifferent and are hydrogen, chlorine, bromine, fluorine, iodine, methyl,ethoxy, methoxy, hydroxy, phosphate, sulfate, nitrate, amino or ethyl; Xis hydrogen, sodium, potassium, or lithium.
 4. A compound of claim 3that inhibits the formation of glucose adducts on albumin.
 5. A compoundof claim 3 that inhibits the formation of glucose adducts on one or moreresidues of albumin as defined by a monoclonal antibody produced by cellline ATCC HB9596.
 6. A compound of claim 3 that is2-[(3-chlorophenyl)amino]phenylacetic acid.
 7. A therapeutic compositioncomprising a compound of claim 1 and a pharmaceutically acceptablecarrier.
 8. A therapeutic composition comprising a compound of claim 2and a pharmaceutically acceptable carrier.
 9. A therapeutic compositioncomprising a compound of claim 3 and a pharmaceutically acceptablecarrier.
 10. A therapeutic composition comprising a compound of claim 4and a pharmaceutically acceptable carrier.
 11. A therapeutic compositioncomprising a compound of claim 5 and a pharmaceutically acceptablecarrier.
 12. A therapeutic composition comprising a compound of claim 6and a pharmaceutically acceptable carrier.
 13. A method of treatingglycation related pathologies in a patient comprising the steps ofadministering to the patient a therapeutically effective amount of atherapeutic composition of claim 7 .
 14. A method for treatingglycation-related pathologies in a patient comprising the steps ofadministering to the patient a therapeutically effective amount of atherapeutic composition of claim 8 .
 15. A method of treating glycationrelated pathologies in a patient comprising the steps of administeringto the patient a therapeutically effective amount of a therapeuticcomposition of claim 9 .
 16. A method of treating glycation relatedpathologies in a patient comprising the steps of administering to thepatient a therapeutically effective amount of a therapeutic compositionof claim 10 .
 17. A method of treating glycation related pathologies ina patient comprising the steps of administering to the patient atherapeutically effective amount of a therapeutic composition of claim11 .
 18. A method of treating glycation related pathologies in a patientcomprising the steps of administering to the patient a therapeuticallyeffective amount of a therapeutic composition of claim 12 .
 19. Aprocess for preparing a compound of claim 3 comprising the steps ofreacting a phenylamine of the formula.

wherein R₂, R₃, R₄, R₅, R₆ may include suitable protection of anyreactive group; with a phenylacetic acid of the formula:

wherein R′₃, R′₄, R′₅, and R′₆ may include suitable protection of anyreactive group and where X is chlorine, bromine, iodine or a sulfonyloxyderivative in the presence of neutralizing agent and copper catalystfollowed by removal of the protecting group, if necessary, to form thedesired product, and, if desired, preparing a salt thereof byconventional means.
 20. A process for preparing a compound of claim 4comprising the steps of reacting a phenylamine of the formula:

wherein: R₂, R₃, R4, R₅, R6 may include suitable protection of anyreactive group; with a phenylacetic acid of the formula:

wherein: R′₃, R′₄, R′₅, R′₆ may include suitable protection of anyreactive group and where X is chlorine, bromine, iodine or a sulfonyloxyderivative in the presence of neutralizing agent and copper catalystfollowed by removal of the protecting group, if necessary, to form thedesired product, and, if desired, preparing a salt thereof byconventional means.
 21. A process for preparing a compound of claim 5comprising the steps of reacting a phenylamine of the formula:

wherein: R′₃, R′₄, R′₅, R′₆ may include suitable protection of anyreactive group; with a phenylacetic acid of the formula:

wherein: R′5 R′₄ R′₅ and R′₆ may include suitable protection of anyreactive group and where X is chlorine, bromine, iodine or a sulfonyloxyderivative in the presence of neutralizing agent and copper catalystfollowed by removal of the protecting group, if necessary, to form thedesired product, and, if desired, preparing a salt thereof byconventional means.
 22. A process for preparing a compound of claim 6comprising the steps of reacting a phenylamine of the formula:

wherein: R′₃, R′₄, R′₅, R′₆ may include suitable protection of anyreactive group; with a phenylacetic acid of the formula:

wherein: R′₅, R′₄ R′₅ and R′₆ may include suitable protection of anyreactive group and where X is chlorine, bromine, iodine or a sulfonyloxyderivative in the presence of neutralizing agent and copper catalystfollowed by removal of the protecting group, if necessary, to form thedesired product, and, if desired, preparing a salt thereof byconventional means.